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ATP-bound form of the D1 AAA domain inhibits an essential function of Cdc48p/p97

Masatoshi Esaki1, Teru Ogura

  • 1Department of Molecular Cell Biology, Institute of Molecular Embryology and Genetics, Kumamoto University, 2-2-1 Honjo, Kumamoto 860-0811, Japan. esaki@kumamoto-u.ac.jp

Biochemistry and Cell Biology = Biochimie Et Biologie Cellulaire
|February 5, 2010
PubMed

Insights

Mutations in Cdc48p/p97, a protein linked to IBMPFD, do not impact its essential functions. However, altering ATPase activity in specific domains, particularly D1, can disrupt cellular processes.

Area of Science:

  • Molecular Biology
  • Cell Biology
  • Biochemistry

Background:

  • Cdc48p/p97 is an essential AAA protein involved in numerous cellular processes.
  • It is implicated in the human disorder inclusion body myopathy with Paget's disease of the bone and frontotemporal dementia (IBMPFD).
  • The protein comprises an N-terminal domain and two ATPase domains (D1 and D2).

Purpose of the Study:

  • To systematically analyze the effects of mutations on the essential functions of yeast Cdc48p/p97 in vivo.
  • To investigate the role of ATPase activities in D1 and D2 domains in protein function.
  • To understand how IBMPFD-related mutations affect Cdc48p/p97 essential functions.

Main Methods:

  • Site-directed mutagenesis
  • Random mutagenesis
  • In vivo functional analysis in yeast

Main Results:

  • IBMPFD-related mutations were found not to affect the essential functions of Cdc48p/p97.
  • Loss of ATPase activity in the D2 domain resulted in a complete loss of protein function in vivo.
  • While D1 ATPase activity alone is not essential, a mutation causing D1 to remain in an ATP-bound state was lethal.
  • Mutagenesis studies indicated that the ATP-bound D1 form disrupts inter-domain interactions, impairing essential functions.

Conclusions:

  • The D2 ATPase activity is critical for Cdc48p/p97 function.
  • The regulation of D1's ATP-bound state is crucial for protein function, suggesting a regulatory role beyond simple ATPase activity.
  • Understanding these domain interactions is key to comprehending Cdc48p/p97's essential cellular roles and its link to IBMPFD.

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